Дисертації з теми "CB7"

In this study, the effects of antagonists to the cannabinoid receptors in MCL cell lines were studied. Results presented in this study show that signalling through cannabinoid receptor with antagonists such as SR141716, SR144528 decreases cell viability but hemopressin when analyzing with XTT. The decrease in cell viability by SR141716 is caused by apoptosis triggered after 5 hours of treatment. The CB1 expression was confirmed in all MCL cell lines tested via western blotting but the expression of CB2 and GPR55 – another receptor to which SR141716 has affinity - was not confirmed due to lack of reliable antibodies. Specific agonist to GPR55 – LPI (l-α-lysophosphatidylinositol) showed different response compared to SR141716 which suggests that the effect seen by SR141716 was not induced through GPR55. The effect induced by CB1/CB2 agonist AEA is shown to be neither through CB1 or CB2 alone but possibly on another receptor yet to be described.

Los receptores de cannabinoides, CB1R y CB2R, que forman parte del sistema endocannabinoide, pertenecen a la superfamilia de receptores acoplados a proteína G (GPCR). El sistema endocannabinoide se encuentra estrechamente relacionado con enfermedades neurodegenerativas que cursan con un proceso de neuroinflamación, como el Parkinson y el Alzheimer. En este proyecto de tesis se ha investigado el papel de los receptores CB1 y CB2 y del heterómero CB1R/CB2R en el contexto de la neuroinflamación. Para ello, se emplearon células N9 inmortalizadas de microglía que expresan los receptores CB1 y CB2, tratadas con LPS e IFN-γ para simular la neuroinflamación, así como tejido y cultivos primarios de animales modelo de la enfermedad de Alzheimer y de la enfermedad de Parkinson. Mediante el uso de técnicas de transferencia de energía y de microscopía confocal se demostró que la formación de heterómeros entre CB1R y CB2R se potencia en condiciones de neuroinflamación. También se constató un aumento de la expresión del receptor CB2 en microglía activada, que afectó a la formación de heterómeros CB1R/CB2R. El análisis de la señalización cannabinoide en células N9 y en los modelos animales demostró que la señalización cannabinoide en microglía activada contrasta con la observada en las células en reposo. Mientras que en reposo se observa un fenómeno de cross-talk negativo entre los receptores CB1 y CB2, en células activadas se observa un cross-talk positivo. Estos resultados convierten al heterómero CB1R/CB2R en una interesante diana terapéutica en procesos neuropatológicos que cursan con neuroinflamación. El sistema endocannabinoide también está implicado en procesos adictivos como el alcoholismo. En adicción alcohólica, una de las estructuras cerebrales más afectadas es el hipocampo. En el segundo apartado de esta tesis doctoral se analizó el efecto del etanol en la señalización de los receptores CB2 y GPR55 (receptor relacionado con el sistema endocannabinoide) y del heterómero CB2R/GPR55, tanto en células HEK-293T transfectadas como en neuronas de hipocampo de rata. Pudo comprobarse que el etanol no altera la heteromerización de los receptores CB2 y GPR55 en células transfectadas. El análisis de la funcionalidad mostró que el etanol no afecta a la señalización por la vía del cAMP del receptor CB2 individual ni del heterómero CB2R/GPR55, ni en células transfectadas ni en cultivos primarios. Este resultado contrasta con la fuerte inhibición en la fosforilación de ERK1/2 observada en ambos modelos celulares, que afectó al receptor CB2R y al heterómero CB2R/GPR55. El análisis de la señalización por calcio del receptor GPR55 en células transfectadas mostró que el etanol potencia la señalización vía calcio de este receptor. Finalmente, mediante técnicas de microscopía confocal, pudo comprobarse que en la corteza frontal de pacientes alcohólicos la heteromerización de los receptores CB2 y GPR55 está potenciada, con un incremento en el número de heterómeros observados con respecto a los individuos sanos. Estos resultados, junto con las evidencias previas de la implicación del sistema endocannabinoide en el alcoholismo, convierten a los receptores CB2 y GPR55 y al heterómero CB2/GPR55 en una interesante diana terapéutica.
CB1 and CB2 receptors are part of the mammalian endocannabinoid system, which is involved in neurologic alterations involving neuroinflammation. Considering these precedents, in this thesis project the main objectives were to demonstrate the implication of cannabinoid receptor heteromers in the regulation of neuroinflammatory processes in activated microglia, in the context of neurodegenerative diseases such as Parkinson’s or Alzheimer’s diseases. Using N9 cells and animal models of Parkinson’s and Alzheimer’s diseases, along with Bioluminescence resonance energy transfer and microscopy techniques, we could demonstrate that the action of cannabinoids on CB1R/CB2R heteromers are potentiated in activated microglia. Using advanced techniques for the study of cell signaling, we were able to prove relevant changes in the signaling of cannabinoid receptors. Whereas in resting cells we could observe negative cross-talk between CB1 and CB2 receptors, in activated microglia this phenomenon changed to a positive cross-talk. Taking in mind these results, and with further investigation, the endocannabinoid system is a promising therapeutic target in neurodegenerative and neuroinflammatory diseases. The endocannabinoid system is also implicated in some addiction processes such as alcoholism. It’s also important to note that in alcoholism the hippocampus is altered. In the second part of this Thesis, we aimed to study the effect of ethanol in the signaling of CB2R, GPR55 (which is related with the endocannabinoid system) and the heteromer CB2R/GPR55. Using transfected HEK-293T cells and primary cell cultures from rat hippocampus, along with Bioluminescence resonance energy transfer and microscopy techniques, we could determine that ethanol did not affect heteromerization. However, cell signaling analysis determined that ethanol inhibits ERK1/2 phosphorylation induced by CB2R and CB2R/GPR55 activation. Calcium signaling analysis of GPR55 resulted in a potentiation of the signal; presumably because of ethanol. We could also study CB2R and GPR55 heteromerization in prefrontal cortex of human alcoholic patients, using proximity ligation assays. Using this approach, we could determine that, compared with age-matched controls, there is an increase in the expression of heteromers. With these interesting results, it is possible to point cannabinoid receptors as therapeutic targets in alcoholism.

Les cannabinoïdes présents dans la marijuana agissent par l'intermédiaire de deux récepteurs, CB1 et CB2, qui sont également activés par des molécules endogènes, les endocannabinoïdes. Les récepteurs CB1, majoritairement exprimés dans le cerveau, relaient les effets psychoactifs du cannabis, mais exercent également de nombreux effets périphériques. Les récepteurs CB2 prédominent dans les cellules du système immunitaire et interviennent notamment dans la régulation de la réponse immune et inflammatoire. Il apparaît aujourd'hui que le système endocannabinoïde joue un rôle crucial au cours des maladies du foie. En effet, le récepteur CB1 participe au développement de l'hypertension portale et de la cardiomyopathie cirrhotique, deux complications de la cirrhose. Il possède des propriétés stéatogènes associées à l'obésité et à la consommation excessive d'alcool. Le récepteur CB2 quant à lui possède des propriétés antifibrogéniques et protège de l'ischémie reperfusion. L'obésité est associée à une réponse inflammatoire qui joue un rôle déterminant dans l'insulino-résistance et la stéatopathie métabolique. Des travaux récents ont montré que le système cannabinoïde favorise le développement de la stéatose par l'intermédiaire des récepteurs CB1. Les récepteurs CB2 interviennent dans la régulation de la réponse immune et inflammatoire. Dans la première partie de ce travail, nous avons donc étudié le rôle des récepteurs CB2 dans le développement de l'obésité, de l'insulino résistance et de la stéatopathie métabolique à l'aide de souris sauvages et invalidées pour le récepteur CB2. Nous avons observé que les souris invalidées pour le récepteur CB2 soumises à un régime hyperlipidique ont une prise de poids significativement plus faible que les souris sauvages consécutivement à une augmentation de l'excrétion fécales des lipides et de l'oxydation des acides gras. Les souris obèses invalidées pour le récepteur CB2 sont plus sensibles à l'insuline et développent une stéatose réduite par rapport aux souris sauvages. Les souris sauvages développent une inflammation importante dans le tissu adipeux viscéral. En revanche, l'induction des cytokines proinflammatoires est significativement plus faible chez les souris CB2-/- exposées au régime hyper lipidique ou chez des souris sauvages obèses traitées avec l'AM630 un antagoniste du récepteur CB2. À l'inverse, l'activation des récepteurs CB2 par un agoniste sélectif conduit à une augmentation de la production de TNFα et de CCL2 dans des explants de tissu adipeux isolés de souris obèses sauvages. Ces résultats constituent la première mise en évidence du rôle des récepteurs CB2 dans le développement de l'obésité, de l'insulino résistance et de la stéatose. Le mécanisme mis en jeu implique probablement un effet proinflammatoire des récepteurs CB2 dans le tissu adipeux. La fibrose est la complication commune de toutes les maladies chroniques du foie et conduit à la cirrhose et à ses complications sévères. Nous avons observé que l'expression de récepteur CB1 est induite dans les zones de fibrose au cours de la cirrhose chez l'homme, notamment dans les cellules fibrogéniques du foie, alors qu'il est peu exprimé dans le foie humain normal. Ces résultats nous ont conduit à évaluer le rôle du récepteur CB1 dans la progression de la fibrose, en étudiant les conséquences de son invalidation génétique et pharmacologique dans trois modèles expérimentaux de physiopathogénie différente, l'administration chronique de tétrachlorure de carbone ou de thioacétamide, ou la ligature de la voie biliaire principale. Dans ces trois modèles, nous avons démontré que l'administration per os d'un antagoniste du récepteur CB1, le rimonabant prévient la fibrogenèse. L'étude des mécanismes impliqués dans les effets antifibrogéniques du rimonabant a révélé que la molécule diminue l'accumulation des cellules fibrogéniques du foie en inhibant leur prolifération. Ces résultats mettent en évidence les propriétés profibrogénique du récepteur CB1 et suggèrent que l'utilisation d'antagonistes du récepteur CB1 pourrait constituer une approche intéressante du traitement de la fibrose.

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El sobrepeso y la obesidad ocasionados por consumo excesivo de alimentos con alto contenido energético es uno de los principales problemas de salud que afectan a la sociedad actualmente. Estas patologías están ligadas a alteraciones del sistema inmunológico que inducen inflamación crónica de bajo grado, que a su vez promueve alteraciones metabólicas como la resistencia a la insulina y el desarrollo de enfermedades crónicas no transmisibles. Una consecuencia de la inflamación crónica es la liberación de moléculas de señalización que inhiben los efectos negativos de la inflamación y regresan al organismo a la homeostasis. Entre ellas, la señalización dependiente de endocanabinoides es esencial para la regulación del metabolismo energético y el funcionamiento del sistema inmunológico, principalmente por la señalización a través de los receptores específicos para endocanabinoides CB1 y CB2. Los receptores CB se localizan en múltiples tipos de células y tejidos, incluyendo el cerebro y los órganos linfoides primarios y secundarios. En años recientes, ha habido un interés mayor en analizar las consecuencias de las modificaciones dietéticas sobre el sistema inmunológico, debido a su papel como el principal sistema de defensa del organismo. Igualmente, ha habido interés en determinar la utilidad de las modificaciones nutricionales para modular funciones fisiológicas relevantes, incluyendo la producción de endocanabinoides y su señalización en diversos tejidos. El objetivo de este estudio fue analizar las diferencias en la expresión de los receptores CB1 y CB2 en el bazo y timo de ratones adultos BALB/c, machos y hembras, suplementados con edulcorantes nutritivos y no nutritivos durante 6 semanas. Nuestros resultados muestran un incremento en la expresión de receptores CB1 en el bazo de ratones hembras suplementados con sucralosa, mientras que los ratones machos del mismo grupo muestran una disminución en la expresión de este receptor, comparados con controles sin suplementación. En contraste, la expresión de CB2 se incrementó en ratones machos suplementados con sacarosa, pero no en las hembras. La expresión de CB1 en el timo mostró un descenso significativo entre los grupos de sucralosa y glucósidos de esteviol en machos, pero no se observaron diferencias en las hembras. Finalmente, la expresión de CB2 en el timo fue más variable, con la sacarosa y sucralosa causando una disminución en la expresión de CB2 en machos y la sacarosa incrementando su expresión en hembras, comparados con los controles. Nuestros resultados muestran que el consumo frecuente de edulcorantes no nutritivos tiene efectos diferenciales dependientes del sexo sobre la expresión de los receptores CB1 y CB2 en el bazo y timo, lo que puede implicar alteraciones en las funciones del sistema inmunológico.
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Orientador: Carlos Amilcar Parada
Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia
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Resumo: A dipirona (metamizol) é um pró-fármaco analgésico utilizado no controle da dor moderada, sendo metabolizada em dois metabolitos bioativos: 4-metil-aminoantipirina (4-MAA) e 4-aminoantipirina (4-AA). O objetivo deste estudo foi investigar a participação de receptores canabinóides periféricos, CB1, CB2 e canais de KATP sobre o efeito anti-hiperalgésico da dipirona, 4-MAA ou 4- AA. Para indução de hiperalgesia, PGE2 (100 ng/pata ) foi administrada localmente na pata traseira de ratos Wistar machos, e o limiar hiperalgésico mecânico foi quantificado por Von- Frey eletrônico, antes e três horas após a injeção. Dipirona, 4-MAA ou 4-AA foram administrados 30 minutos antes do Von Frey. Os antagonistas seletivos do receptor CB1 (AM251), CB2 (AM630) e glibenclamida, um bloqueador KATP (80 ug) ou ODQ um inibidor de cGMP (32 ?g) foram administrados 30 minutos antes da Dipirona, 4-MAA ou 4 -AA. O ODN-antisense para reduzir a expressão do receptor CB1 (30 ?g) foi administrado por via intratecal, uma vez por dia durante quatro dias consecutivos. A hiperalgesia mecânica induzida pela PGE2 foi reduzida pela dipirona, 4-MAA, e 4-AA de maneira dose-dependente. AM251 ou ODN-antisense contra o receptor neuronal CB1, mas não AM630, reduziu o efeito anti-hiperalgésico mediado por 4-AA, mas não da dipirona ou 4-MAA. Por outro lado, o efeito anti-hiperalgésico da dipirona, ou 4-MAA foi revertido por glibenclamida ou ODQ. Os resultados sugerem que a ativação de receptores neuronal CB1, mas não do receptor CB2, no tecido periférico esteja envolvido no efeito anti-hiperalgésico do metabólito 4-AA. Além disso, a dipirona e 4-MAA possui um efeito anti-hiperalgesico dependente de cGMP e consequente abertura KATP
Abstract: Dipyrone (metamizole) is an analgesic pro-drug used to control moderate pain. It is metabolized in two bioactive metabolites: 4-methylaminoantipyrine (4-MAA) and 4-aminoantipyrine (4-AA). The aim of this study was to investigate the participation of peripheral CB1 and CB2 cannabinoid receptors activation on the anti-hyperalgesic effect of Dypirone, 4-MAA or 4-AA. For induction of hyperalgesia, PGE2 (100 ng) was locally administrated in hindpaw of male Wistar rats, and the mechanical nociceptive threshold was quantified by electronic von-Frey, before and 3 hours after its injection. Dypirone, 4-MAA or 4-AA was administrated 30 minutes before the von-Frey test. The selective CB1 receptor antagonist AM251, CB2 receptor antagonist AM630, cGMP inhibitor ODQ (32 ?g) or KATP blocker glibenclamide (80 ?g) was administrated 30 minutes before Dypirone, 4-MAA or 4-AA. The antisense-ODN against CB1 receptor expression (30 ?g) was intrathecally administrated once a day during four consecutive days. PGE2-induced mechanical hyperalgesia was inhibited by dypirone, 4-MAA, and 4-AA in a dose-response manner. AM251 or ODN anti-sense against neuronal CB1 receptor, but not AM630, reversed the antihyperalgesic effect mediated by 4-AA, but not by dypirone or 4-MAA. On the other hand, the anti-hyperalgesic effect of dypirone or 4-MAA was reversed by Glibenclamide or ODQ. These results suggest that the activation of neuronal CB1, but not CB2 receptor, in the peripheral tissue is involved in the anti-hyperalgesic effect of 4-aminoantipyrine. In addition, 4- methylaminontipyrine mediates anti-hyperalgesic effect by the cGMP activation and the KATP opening
Mestrado
Fisiologia
Mestre em Biologia Funcional e Molecular

Los receptores cannabinoides CB1R y CB2R son GPCRs que forman parte del sistema endocannabinoide y son considerados dianas potenciales para numerosas enfermedades. Su compleja farmacología junto con la naturaleza lipofílica de sus ligandos perjudica la traslacionalidad de la investigación centrada en el sistema endocannabinoide. Para comprender mejor la farmacología de CB2R se puso a punto una técnica homogénea y no radioactiva para estudiar la unión de ligando en célula viva, gracias a la cual se identificó un segundo centro de afinidad en CB2R, y cambios conformacionales en el receptor cuando éste se coexpresa con GPR55, un GPCR huérfano que une cannabinoides. Los GPCRs pueden formar heterooligómeros, con propiedades farmacológicas diferentes a las de los receptores individuales. Dado que existen complejos CB1R-CB2R y CB2R-GPR55, se investigó si CB1R y GPR55 podían formar complejos heteroméricos. Se confirmó esta posibilidad en un sistema de expresión heterólogo y en muestras de sistema nervioso central, donde se detectó la huella del heterómero. Se postula que en microglía la activación de los receptores de cannabinoides produce un efecto neuroprotector. El GPCR huérfano GPR18, que une cannabinoides y comparte con CB2R un papel regulador de la respuesta inmune, se expresa en microglía. Dado que se ha sugerido un posible cross-talk entre CB2R y GPR18, se estudió si GPR18 era capaz de interaccionar con CB1 y CB2. Se observó que GPR18 no es capaz de formar complejos heteroméricos con CB1R, pero si con CB2, tanto en un sistema heterólogo como en microglía, y en ambos casos se detectó la huella del heterómero. En microglía activada y en cultivos primarios de ratones modelo de la enfermedad de Alzheimer la expresión de CB2R-GPR18 se encuentra aumentada. Dado el papel de CB2R en microglía, se estudió la presencia y la función de los complejos CB1R-CB2R en microglía en reposo y activada. La expresión aumentada de complejos CB1R-CB2R y su señalización en microglía activada indican que la función de los cannabinoides es la de mantener la microglía en reposo, pero desencadenar una señalización robusta cuando la microglía se activa. En cerebro de animales modelo de enfermedades neurodegenerativas (Parkinson y Alzheimer), donde se encontraron marcadores de microglía activada, se observaron resultados similares a los encontrados en una línea de microglía tratada con lipopolisácarido e interferón gamma. Ello sugiere que CB1R, CB2R y CB1R-CB2R en microglía activada tienen potencial como dianas en el tratamiento de enfermedades neurodegenerativas que cursan con neuroinflamación. Los receptores CB2R y GPR55 tienen un relevante papel en mecanismos de respuesta a estrés, ansiedad y depresión. Considerando la estrecha relación entre las enfermedades neuropsiquiátricas y el suicidio, se evaluó si la expresión de los complejos CB2R-GPR55 está alterada en suicidas. Se detectó la presencia de heterómeros CB2-GPR55 en corteza prefrontal, y un aumento en la expresión de estos heterómeros en corteza prefrontal de muestras de suicidas. Para investigar el potencial terapéutico de los fitocannabinoides es necesario conocer mejor su relación con CB1R y CB2R, ya que el mecanismo de acción del cannabidiol y el cannabigerol (dos importantes componentes no psicotrópicos de la planta del cannabis) sobre estos receptores es aún confuso. Se observó que a concentraciones nanomolares el cannabidiol es capaz de modular de manera negativa la tanto la afinidad de la unión de los agonistas a CB2R como la señalización mediada por este receptor, por lo que se postula que el cannabidiol es un modulador alostérico de CB2R. En cambio, cannabigerol parece unirse al centro ortostérico de CB1R y CB2R, sobre los que actúa como un agonista con selectividad funcional, y modula la señalización de los cannabinoides sobre los heterómeros CB1R-CB2R.
Cannabinoid CB1 (CB1R) and CB2 (CB2R) receptors are GPCRs of the mammalian endocannabinoid system.Their complex pharmacology is delaying the translational success of medications targeting the endocannabinoid system. To better understand CB2R pharmacology, a novel homogeneous technique was developed to study ligand binding to CB2R in living cells, which disclosed a second affinity state of CB2R and conformational changes when CB2R interacts with another GPCR. To know how cannabinoid signaling is affected by receptor-receptor interactions, we studied whether CB1R and CB2R interact with two orphan GPCRs, namely GPR55 and GPR18. CB1R-GPR55 and CB2R-GPR18 heteromers were detected both in transfected cells and in the central nervous system, and their fingerprint was also revealed. Given the neuroprotective role of CB2R activation to prevent neuroinflammation, the presence of CB1R-CB2R and CB2R-GPR18 heteromers was studied in both resting and activated microglia. The higher number of complexes and the differential signalling found both in activated microglia and in the brain of animal models of neurodegenerative diseases, indicate that CB1R-CB2R and CB2R-GPR18 heteromers have a role in neuroprotection. Other pathologies where cannabinoid receptors have shown an important role are anxiety and depression disorders, which are related to suicide. The presence of the previously described CB2R-GPR55 heteromer was detected in human prefrontal cortex, and a higher amount of heteroreceptor complexes was found in samples from suicide victims, indicating that CB2R-GPR55 heteromers could have a role in depression. To further exploit the therapeutic potential of the cannabinoids, it is also important to understand how phytocannabinoids, such as cannabidiol and cannabigerol, two compounds found in the cannabis plant and which lack psychoactive effects, interact with CB1R and CB2R. Nanomolar concentrations of cannabidiol modulated in a negative way the affinity of the binding of agonists to CB2R as well as the signaling mediated by this receptor. Thus, it is postulated that cannabidiol is an allosteric modulator of CB2R. In contrast, cannabigerol appears to bind to the orthosteric center of CB1R and CB2R, where it acts as an agonist with functional selectivity, and modulates the signaling of the cannabinoids on CB1R-CB2R heteromers.

Tesis de Maestría
Los receptores CB1 y CB2 del sistema endocanabinoide tienen funciones importantes en el hígado y el páncreas, ya que participan en funciones esenciales como la regulación del metabolismo celular, la liberación de insulina y la homeostasis de la glucosa. En general, las dietas de alta energía promueven la activación de los receptores CB, que cuando se estimulan crónicamente, se asocian con anomalías metabólicas e inflamación en condiciones como la obesidad y la diabetes. La ingestión de edulcorantes no nutritivos se ha expandido en poblaciones de todo el mundo con el propósito de reducir el contenido energético de la dieta y para el control de enfermedades crónicas asociadas con alteraciones del metabolismo energético; sin embargo, se desconocen los posibles efectos de estos compuestos sobre la regulación del sistema endocanabinoide a nivel hepático y pancreático. El objeto de estudio fue evaluar la presencia de cambios en la expresión de los receptores CB1 y CB2 en páncreas e hígado de animales experimentales a quienes se les suministraron edulcorantes no nutritivos comerciales (glucósidos de esteviol o sucralosa) en un período de 6 semanas. Después de la suplementación, se analizó la expresión de los receptores CB1 y CB2 de ambos tejidos mediante la técnica de western blot e inmunofluorescencia. Los resultados muestran reducciones significativas en la expresión de CB1 y CB2 en hígado, particularmente en animales machos suplementados con glucósidos de esteviol, con tendencias hacia una expresión reducida de CB1 en páncreas en el mismo grupo. Por el contrario, hubo una tendencia hacia una mayor expresión de CB1 hepático en animales hembra suplementados con sacarosa, sucralosa o glucósidos de esteviol. Estos datos apuntan a que la ingestión frecuente de edulcorantes no nutritivos específicos promueve alteraciones dependientes del sexo en la expresión de los receptores CB en el páncreas y el hígado in vivo.

Orientadores : Marcos Garcia Costa, Luiz Candido S. Dias
Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia
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Mestrado
Imunologia
Mestre em Ciências Biológicas

CBI Betonginstitutet är idag i en expansionsfas med en strävan att värna om den globalahållbarheten som medför större förväntningar på företaget och ökade krav från samhället somi världen. För att konkurrera i näringslivet medför detta en förändring i verksamheten och iverksamhetens processer. Detta ger ökad medvetenhet i helhet hos medarbetaren för attsamverka i organisationen och internationellt. För att kunna förbättras krävs det gemensamförståelse från samtliga medarbetare oavsett titel, där de gemensamt utvecklas kringorganisationen mål och visioner.Syftet med examensarbetet är att hjälpa CBI med sammanställning av deras huvudprocesser iledningssystemet samt en enkel överskådlig processkarta som smidigt kan integreras i SP:sverksamhet.Rapporten ger en redogörelse hur teori och praktiskt genomförande skiljs åt. Material tillexamensrapport, fick fram genom att studera CBI ledningssystem och genom litteraturstudiersamt avslutande intervjuer med medarbetare.Resultatet utgår från medarbetarnas beskrivning av deras arbetsmoment, därifrån skapades enprocesskarta för huvudprocesser och delprocesser som grund till CBI ledningssystem. Visaräven hur huvudprocessen provning & kontrolls, ackrediterad verksamhet, delprocesser harminskat från nio stycken till fem stycken delprocesser.I framtida arbeten med förbättring och fortsatt utveckling av processer för att få en smidigvärdeskapande kedja av flöde och för medarbetare att se helheten i organisation. Därledningens mål och visioner speglas ner i verksamheten och utvecklandet av processer.

Program: Industriell ekonomi - arbetsorganisation och ledarskap

Program: Industriell ekonomi - affärsingenjör

The full abstract for this thesis is available in the body of the thesis, and will be available when the embargo expires.
Medicine, Faculty of
Obstetrics and Gynaecology, Department of
Graduate

Este trabalho tem como objetivo rever as relações existentes entre os valores obtidos dos ensaios mini-CBR e CBR, a fim de tornar possível a utilização dos resultados de ensaios de mini-CBR no dimensionamento de pavimentos flexíveis, uma vez que este ensaio é mais prático, exige menor quantidade de amostra, é mais rápido para ser executado e exige menor esforço físico na sua realização, sendo, portanto, menos dispendioso que o ensaio de CBR. No desenvolvimento dos ensaios, os resultados indicavam que essas relações não se mostravam adequadas para o universo em estudo, pois quando tentava-se relacionar, graficamente, os resultados dos ensaios tradicionais de CBR com os do mini-CBR, não se obtinham retas como era de se esperar e sim uma nuvem de pontos, indicando não haver correlação entre os dois índices. Desta forma, tentou-se desenvolver novas relações, nos moldes do que fora feito originalmente por Nogami (1972), e correlacionar novamente os valores de carga do ensaio de CBR, para as penetrações de 2,5 e 5,0 mm, com os valores de carga desenvolvidos nos ensaios padronizados de mini-CBR, para as penetrações de 2,0 e 2,5 mm, respectivamente, nas energias de compactação normal e intermediária. Nesta pesquisa são abordados os fundamentos, a descrição e algumas pesquisas realizadas com os ensaios de CBR e de mini-CBR. Foram utilizadas nesta pesquisa 62 amostras de solos do município de São Carlos - SP, as quais foram submetidas ao ensaio de CBR, nas energias de compactação normal e intermediária, e os resultados foram comparados com os dos ensaios de mini-CBR. As amostras foram escolhidas considerando-se a proporcionalidade entre as áreas de ocorrência das classes pedológicas no município de São Carlos. Por fim, apresentam-se as tentativas feitas para aproveitar ao máximo os resultados obtidos nos ensaios de CBR e de mini-CBR.
The objective of this work is to review the relation among the results of mini-CBR and CBR tests. This will make possible the use of mini-CBR tests results in flexible pavements design, once this test is more practical, it demands smaller amount of samples, it execution is faster and it demands less physical effort, being, therefore, less expensive than CBR tests. In the development this tests, the results indicated that those relations not shown appropriate for the universe in study, therefore when it tried to relation, graphically, the results of the traditional CBR test with the one of the mini-CBR, not obtained straight line as it was of waiting and yes a cloud of points, indicating not to be correlation among the two indexes. This form, tried to develop new relations, in the molds of what had been done originally by Nogami (1972), and to correlate the values of load CBR test, for the penetrations of 2,5 and 5,0 mm, with the load values developed in the standardized mini-CBR test, for the penetrations of 2,0 and 2,5 mm, respectively, in normal and intermediate compactation energies. In this tests are foundations, the description and some researches realized are approached with the CBR and mini-CBR tests. In this tests will used, 62 soils samples from São Carlos - SP, will be submitted to CBR test, in normal and intermediate compactation energies, and the results will be compared with those ones from mini-CBR test. The samples were chosen considering theproportionality among pedologic class occurrence areas in São Carlos. Finally, to show up the attempts done to utilize of to the maximum the results obtained in the CBR and mini-CBR tests.

This study investigated the prevalence of voluntary self-practice of Cognitive Behaviour Therapy (CBT) by accredited CBT practitioners, and explored the ability of the Theory of Planned Behaviour (TPB) to predict intentions to engage in CBT self-practice. A TPB questionnaire was constructed by implementing the protocol devised by Francis et al. (2004), with content specific items generated from an elicitation exercise to obtain salient beliefs regarding CBT self-practice. A sample of 177 accredited CBT practitioners completed an online survey which included demographic items and the TPB questionnaire. It was found that 46.4% of participants intended to self-practice CBT more than once a week over the following month. While the TPB predicted intentions to self-practice CBT (R2 = .445, p < .001), structural equation modeling revealed that the theory of reasoned action (R2 = .473, p < .001) was the best model fit of the observed data (RMSEA = .00, CFI = 1.00). It was concluded that a substantial proportion of CBT practitioners intended to regularly engage in CBT self-practice, with intentions predicted by subjective normative beliefs and attitudes towards the behaviour. Implications are discussed, with calls for the study to be replicated. Potential future research is considered, with suggestions to explore the role of CBT self-practice to promote therapist wellbeing.

En este informe se ha realizado una estimación del valor económico de la empresa CBC Peruana S.A.C., al 31 de diciembre de 2018, aplicando como método para su valoración el flujo de caja libre descontado (FCFF). CBC Peruana es una empresa de reciente creación (2015) dedicada a la fabricación, distribución y comercialización de bebidas no alcohólicas. Forma parte del grupo guatemalteco The Central America Bottling Corporation (CBC) y, en el Perú, es la embotelladora y distribuidora exclusiva de PepsiCo; a nivel nacional cuenta con dos plantas embotelladoras y su distribución llega a más de 200 mil puntos de venta. En el Perú, la industria de bebidas no alcohólicas ha experimentado un rápido crecimiento en las categorías de bebidas energéticas, isotónicas, agua embotellada y still drinks; sin embargo, en los últimos años, el crecimiento de las bebidas carbonatadas se ha visto afectado por la creciente tendencia del mercado al consumo de bebidas saludables. El crecimiento de este sector se encuentra fuertemente correlacionado con el PIB y la demanda interna, por lo que se proyecta un mayor crecimiento de este mercado, especialmente en las categorías antes mencionadas. El objetivo de CBC Peruana está orientado a lograr una mayor penetración de mercado a través de la utilización de estrategias de diferenciación y liderazgo en costos. Actualmente, sus principales competidores son Arca Continental y el Grupo Aje. Para la valoración han sido proyectados los flujos para un periodo de 10 años (2019-2028).

Validazione del CBA-Young Form (CBA-Y) Project of research "CBA young form: validation of a wide spectrum questionnaire for screening of the protective and risk factors linked with psychological disease of young people" Introduction CBA group build up a psychological tool thought to the screening of young people: CBA young form. The questionnaire born to cover an Italian lack concerned the psychological tools for people aged within 14 and 25 years old (Sanavio, et al., 1992; Michelin & Sica, 1996). The aim of CBA young form is evaluating the typical trouble of adolescence in Italy. The main goal of the questionnaire is the evaluation of three things: psychological wellness, the probability whom the subject could have some kinds of dangerous behaviors and the subject's skills against negative events in life. The diagnosis of the state of well-being and psychological adjustment of the subject is a priority goal for any instrument of medical, also referring to an age evolving as that of adolescence or youth, it becomes essential to collect elements that can help the specialist to provide for "risky behavior" on the part of the subject. Alcohol, drugs, AIDS, car accidents, vandalism, etc.. seriously put at risk the lives of many young people and invariably mark the future adulthood. Finally, it is necessary to be able to estimate the presence of so-called "coping strategies" coping strategies can be conceptualized as psychological resources that help the person to develop and address the mutavoli situations she encounters in the course of their lives (Goldfried and Davison , 1976; Petter, 1999). In fact, insufficient acquisition of these strategies results in difficulties and problems in interpersonal behavior and social life of the individual. On the other hand, realistic expectations of self-efficacy, the ability to restructure erroneous beliefs, a sense of self-confidence, problem solving skills, etc.., Can be considered as real factors "protective" of the psychological health of the person and more generally an indicator of well-being and psychosocial adjustment. Description of the CBA-Y The CBA-Y in its final form consists of 500 items by filling in the form checklist. Who fills in the questionnaire flows individual item and mark in the box corresponding to those judges can adapt to your situation. This mode of response is generally considered to be less invasive because it allows the omission of the item rather than forcing the individual to give an answer. Compilation time is around 30 minutes. Protocol CBA-Y are obtained across a range of information about individual items marked dall'esaminato. In addition there is the structuring of responses within two control scales (Autoapertura, Emotional State), two scales of well-being (well-being, psychosocial adjustment) and 7 scales of discomfort (Use of Drugs and Unusual Experiences, psychopathological risk, Love the risk and search of excitement, anxiety and emotional instability, Characteristics and antisocial behavior, irritability, aggressiveness and hostility Events psychophysiological). It also calculates the number of items that were used to describe as additional information for the psychologist. At the same time it was realized the short form of the same questionnaire, the Youth Welfare CBA (CBA-BG). The short form differs from the baseline questionnaire for the reduction in the number of items that have been brought to 200. The stairs that are calculated has been eliminated on the emotional state portandone the total number to 10. Objectives The overall objective of the research is to validate the CBA Young form. The validation process is divided into different times with different priorities. 1. Investigate the main psychometric characteristics of the questionnaire. We intend to evaluate the factor structure of the questionnaire, the distribution of scores in the scales and their reliability, possible gender differences in the responses and any differences related to age groups. 2. Convergent and discriminant validity: comparison of the instrument with other psychological screening questionnaires in charge of the youth population. 3. Criterion validity: to observe the pattern of response to the questionnaire populations characterized by the presence of specific symptoms in the field of psychology. 4. Measure the predictive power of the questionnaire to the onset of psychopathological symptoms in young people. Participants A large representative sample of the normal population (about 2000 participants). Subsamples pathological to test the response of the questionnaire when administered to individuals critical. A sample external to that used for the collection of normative data to test the external validity of the questionnaire. Materials and methods - Informed Consent Form - Folder registry (socio demographic information, medical history and related to pharmacological treatment in progress) - Medical Record (information on the diagnosis and course of the disease) - Young form CBA (CBA-Y) - CBA youth welfare (CBA-BG) - Minnesota Multiphasic Personality Inventory version adolescents (Butcher & Williams, 1996). The number of participants will take part in the compilation of one or more questionnaires. Partecipanti Un vasto campione rappresentativo della popolazione normale (circa 2000 partecipanti). Sottocampioni patologici per verificare la risposta del questionario se somministrato a individui critici. Un campione esterno a quello utilizzato per la raccolta dei dati normativi per testare la validità esterna del questionario. Materiali e metodo - Modulo di consenso informato - Cartella anagrafica (informazioni socio anagrafiche, anamnestiche e relative al trattamento farmacologico in corso) - Cartella clinica (informazioni relative alla diagnosi e al decorso della malattia) - CBA young form (CBA-Y) - CBA benessere giovanile (CBA-BG) - Minnesota Multiphasic Personality Inventory versione adolescenti (Butcher & Williams, 1996). I diversi partecipanti dovranno partecipare alla compilazione di uno o più questionari. Study 1: analysis of the differences between the response mode "checklist" and how to respond "true or false". Goal. The objectives of this work are twofold: to assess whether the checklist mode with respect to the true / false influence scores and highlight any differences between paper and electronic administration. Method. 100 people completed the questionnaire in response mode checklist and true / false in order counterbalanced. 50 people have made the two compilations in the electronic version and 50 in the paper version. To evaluate the differences between the two modes of response was used the McNemar test on the 16 sections of the questionnaire and the analysis of variance on the scores obtained in 15 steps. To evaluate the differences between paper and electronic administration has been used analysis of variance on the scores obtained in 15 steps. Results. The McNemar test showed that the number of items marked "true" is greater than the number of items selected in the checklist in all sections of the questionnaire included the "status anxiety." The ANOVA showed that the mean scores in the scales of the questionnaire are higher in answer mode "true / false" with respect to the checklist. There were no differences between the paper and the electronic administration. Study 2: Validation of the CBA-Y. Goal. Validate the questionnaire CBA-Y on the Italian population. Method. For the validation of CBA-Y were recruited 1732 children, 842 males (48.6%) and 890 females (51.4%) aged between 14 and 25 years. It 'a community group, recruited on a voluntary basis in schools, workplaces and other settings supposedly' normal '; 92% resides in Northern Italy. Everyone was instructed about how to complete the questionnaire and freely accepted to take part in the compilation informed consent. During compilation that could be both individual and group psychologist has always been available for any questions related to the task. The collected data were then analyzed to derive threshold values belonging to each scale of the questionnaire divided by gender and age groups. The reliability of the scales was investigated by Cronbach's alpha index (Cohen, 1988). The convergent validity (Campbell & Fiske, 1959) of the questionnaire was analyzed by correlating the questionnaire with the MMPI version adolescents. Results. The questionnaire has revealed a good degree of reliability and internal validity. The relationship with the MMPI is detected consistent with what is present in the literature. Study 3: Application of the CBA-Y to a clinical population. Goal. Investigate whether the CBA-Y discriminate between normal and pathological population. Method. Looking took part in 34 patients with a clinical diagnosis on axis I and II. The average age of the entire sample was 20.5 years (range 14-26; Ds 3.66). Within the sample children in treatment were divided into five subgroups based on the principal diagnosis: anxiety, mood, personality, and substance use disorders of adolescence. The delivery took place at the services of Pediatrics, or at home on the experimenter sending the psychotherapist. To investigate the differences between the normative sample and the clinical sample was used the test t-student. Results. Analysis of data shows a good discriminating power of the CBA-Y to distinguish the normal population from the clinic. Study 4: Differences between normative data and a sample of French students. Goal. This study aims to investigate how a population different from the Italian answer to the CBA-BG to investigate aspects of external validity. Method. The Italian version of the questionnaire (the original version) was translated into French and then reported in the original language (back Traslation) in agreement with the suggestion made in the article by Brislin 1970. The questionnaire was administered in French in the Faculty of Psychology of Nanterre (Paris Ouest) during breaks between classes. Each student completed the questionnaire individually and has previously been instructed to ask any questions the examiner. Surveyed 109 students (50 males, 45.9%) French aged between 18 and 24 years. The average age of the children was 21.98 years (SD = 2.44) compared to women was 20.81 years (sd = 1.51). The age between the two genders differed significantly (F (1,107) = 8.35, p <0.01). All respondents were students of the psychology faculty of Nanterre. There is no statistically significant difference between the number of boys and girls (χ2 (1) = 0,743, p = 0,389). Analysis: to confirm the factor structure within the French sample were used for structural equation modeling in their simplest form, the confirmatory factor analysis (CFA). Because of the scarcity of the sample each scale of the CBA-BG was evaluated individually in order to reduce the complexity of the model. To investigate whether there were differences between the responses of the French champion than a sub-sample extracted from matched normative data was used univariate analysis of variance using factors such as gender and group (French and Italian). The results of the analysis are considered the interaction between the factors and the main effect of group. The first explicit the presence of differences between gender differences in the two groups while the second investigates the possible difference between the groups. The main effect of gender is irrelevant for the purposes of this research. Results. The analysis on the French champion has confirmed the validity of the questionnaire for seven of the ten scales analyzed. The only partial adherence of the model to the original data can be attributed to the nature of the scales of CBA-BG. They were not obtained through statistical procedures but are designed in theory to investigate the different areas in preparation of the instrument were considered useful for clinical practice. The two samples tested were equivalent in all important aspects. In addition to pairing by gender, age, education and professional status they did not differ even for the conditions of administration. A variable difficult to control in the analysis that often do not pay particular attention during data collection. The analysis of the scores obtained in the two samples did not reveal substantial differences between the scales of the questionnaire. Only the scale of well-being and the number of items used to describe show a score significantly lower than that in the sample French Italian. We can conclude by saying that the results of this study to generalize the data obtained in the first phase of validation of the questionnaire are broadly satisfactory.
Progetto di ricerca “CBA forma Giovanile: validazione di una questionario ad ampio spettro per lo screening dei fattori protettivi e predisponenti al disagio psicologico nella popolazione giovanile”. Quadro di riferimento Nello sviluppo delle ricerche del gruppo CBA è stato ideato uno strumento di indagine clinica per la popolazione giovanile: il CBA forma giovani. Il questionario nasce per colmare una lacuna che esiste in Italia per quanto riguarda gli strumenti di assessment e ricerca che siano adatti per persone tra i 14 e 25 anni (Sanavio, et al., 1992; Michelin & Sica, 1996). Il CBA forma Giovani ha lo scopo di valutare le problematiche proprie della tarda adolescenza e della giovinezza nel contesto italiano. Nella costruzione del questionario si sono ritenuti prioritari tre obiettivi: valutare il benessere psicologico e l’adattamento psicosociale del soggetto, valutare la probabilità che si verifichino condotte che potrebbero ledere il soggetto stesso o altre persone e valutare le risorse del soggetto per far fronte ad eventi avversi nella propria vita. La diagnosi dello stato di benessere e adattamento psicologico del soggetto è uno scopo prioritario per qualsiasi strumento di carattere clinico; inoltre, riferendosi ad una fascia d’età in piena evoluzione come quella della adolescenza o della giovinezza, diventa fondamentale poter raccogliere degli elementi che possano aiutare lo specialista a prevedere dei “comportamenti a rischio” da parte del soggetto. Alcool, droga, AIDS, incidenti stradali, teppismo, ecc. mettono seriamente a repentaglio la vita di moltissimi giovani o segnano invariabilmente la futura età adulta. Infine è necessario poter fare una stima della presenza delle cosiddette “strategie di coping”; le strategie di coping possono essere concettualizzate come delle risorse psicologiche che aiutano la persona ad elaborare ed affrontare le mutavoli situazioni che incontra nel corso della propria vita (Goldfried e Davison, 1976; Petter, 1999). Infatti, l’insufficiente acquisizione di tali strategie si traduce in difficoltà e problemi nel comportamento interpersonale e nella vita sociale dell’individuo. Per contro, aspettative realistiche di auto efficacia, la capacità di ristrutturare convinzioni erronee, un senso di fiducia nelle proprie capacità, abilità di problem solving, ecc., possono essere considerati dei veri e propri fattori “protettivi” della salute psicologica della persona e più in generale un indicatore di benessere e adattamento psicosociale. Descrizione del CBA-Y Il CBA-Y nella sua forma definitiva è composto da 500 item da compilare in forma checklist. Chi compila il questionario scorre i singoli item e segna la casella corrispondente a quelli che giudica possano adattarsi alla sua situazione. Questa modalità di risposta è giudicata in generale come meno invasiva perché consente l'omissione dell'item anziché forzare l'individuo a dare una risposta. Il tempo di compilazione si aggira attorno ai 30 minuti. Dal protocollo CBA-Y si ricavano tutta una serie di informazioni inerenti i singoli item marcati dall'esaminato. Inoltre è prevista la strutturazione delle riposte all'interno di 2 scale di controllo (Autoapertura, Stato Emotivo), 2 scale di benessere (Benessere, Adattamento psicosociale) e 7 scale di disagio (Uso di Droghe ed Esperienze Inusuali, Rischio Psicopatologico, Amore per il rischio e ricerca di sensazioni forti, Ansia e Instabilità Emotiva, Caratteristiche e comportamenti antisociali, irritabilità Aggressività Ostilità e Manifestazioni Psicofisiologiche). Viene inoltre calcolato il numero di item che sono stati usati per descriversi come ulteriore informazione per lo psicologo. Parallelamente è stata realizzata la forma breve dello stesso questionario, il CBA Benessere Giovanile (CBA-BG). La forma breve si differenzia dal questionario di base per la riduzione nel numero degli item che sono stati portati a 200. Dalle scale che vengono calcolate è stata eliminata quella relativa allo stato emotivo portandone il numero totale a 10. Obiettivi L’obiettivo globale della ricerca è validare il CBA forma Giovani. Il processo di validazione si articola in diversi momenti con differenti priorità. 1. Indagare le principali caratteristiche psicometriche del questionario. Si intende valutare la struttura fattoriale del questionario, la distribuzione dei punteggi nelle scale e la loro attendibilità, eventuali differenze di genere nelle risposte e eventuali differenze legate alle classi di età. 2. Validità convergente e discriminante: confronto dello strumento con altri questionari preposti allo screening psicologico della popolazione giovanile. 3. Validità di criterio: osservare i pattern di risposta al questionario di popolazioni caratterizzate dalla presenza di sintomatologie specifiche in ambito psicologico. 4. Misurare il potere predittivo del questionario verso l’insorgenza di sintomi psicopatologici nella popolazione giovanile. Partecipanti Un vasto campione rappresentativo della popolazione normale (circa 2000 partecipanti). Sottocampioni patologici per verificare la risposta del questionario se somministrato a individui critici. Un campione esterno a quello utilizzato per la raccolta dei dati normativi per testare la validità esterna del questionario. Materiali e metodo - Modulo di consenso informato - Cartella anagrafica (informazioni socio anagrafiche, anamnestiche e relative al trattamento farmacologico in corso) - Cartella clinica (informazioni relative alla diagnosi e al decorso della malattia) - CBA young form (CBA-Y) - CBA benessere giovanile (CBA-BG) - Minnesota Multiphasic Personality Inventory versione adolescenti (Butcher & Williams, 1996). I diversi partecipanti dovranno partecipare alla compilazione di uno o più questionari. Studio 1: analisi delle differenze tra la modalità di risposta "cheklist" e la modalità di risposta "vero o falso". Obiettivo. Gli obiettivi del presente lavoro sono due: valutare se la modalità checklist rispetto al vero/falso influenza i punteggi ed evidenziare eventuali differenze tra somministrazione cartacea ed elettronica. Metodo. 100 persone hanno compilato il questionario in modalità di risposta checklist e vero/falso in ordine controbilanciato. 50 persone hanno effettuato le due compilazioni in versione elettronica e 50 in versione cartacea. Per valutare le differenze tra le due modalità di risposta è stato utilizzato il test di McNemar sulle 16 sezioni del questionario e l’analisi della varianza sui punteggi ottenuti nelle 15 scale. Per valutare le differenze tra somministrazione cartacea ed elettronica è stata utilizzata l’analisi della varianza sui punteggi ottenuti nelle 15 scale. Risultati. Dal test di McNemar è emerso che il numero di item siglati “veri” è maggiore del numero di item selezionati nella checklist in tutte le sezioni del questionario esclusa la sezione “ansia di stato”. L’ANOVA ha evidenziato che i punteggi medi nelle scale del questionario sono più alti nella modalità di risposta “vero/falso” rispetto alla checklist. Non si sono riscontrate differenze tra la somministrazione cartacea e quella elettronica. Studio 2: validazione del CBA-Y. Obiettivo. Validare il questionario CBA-Y sulla popolazione italiana. Metodo. Per la validazione del CBA-Y sono stati reclutati 1732 ragazzi, 842 maschi (48,6%) e 890 femmine (51,4%) di età compresa tra 14 e 25 anni. E’ un gruppo di comunità, reclutato su base volontaria in scuole, luoghi di lavoro e altri contesti presumibilmente ‘normali’; il 92% risiede nel Nord-Italia. Ogni individuo è stato istruito circa la modalità di compilazione del questionario e ha accettato liberamente di prendere parte alla compilazione previo consenso informato. Durante la compilazione che poteva essere sia individuale sia di gruppo uno psicologo è sempre rimasto disponibile per gli eventuali chiarimenti inerenti il compito. I dati raccolti sono stati poi analizzati per ricavare i valori di soglia appartenenti a ciascuna scala del questionario suddivisi per genere e fasce d'età. L'affidabilità delle scale è stata indagata attraverso l'indice alfa di Cronbach (Cohen, 1988). La validità convergente (Campbell & Fiske, 1959) del questionario è stata analizzata mettendo in relazione il questionario con l'MMPI versione adolescenti. Risultati. Il questionario ha rivelato un buon grado di affidabilità e di validità interna. La relazione con l'MMPI si rileva coerente con quanto presente in letteratura. Studio 3: Applicazione del CBA-Y ad una popolazione clinica. Obiettivo. Indagare se il CBA-Y discrimina tra la popolazione normale e patologica. Metodo. Alla ricerca hanno preso parte 34 pazienti con diagnosi clinica in asse I e II. L'età media dell'intero campione è di 20,5 anni (range 14-26; Ds 3,66). All'interno del campione i ragazzi in trattamento sono stati divisi in 5 sottogruppi in base alla diagnosi principale: ansia, umore, personalità, disturbi dell'adolescenza e uso di sostanze. La somministrazione è avvenuta presso i servizi di Neuropsichiatria Infantile, o a domicilio dello sperimentatore su invio dello psicoterapeuta. Per indagare le differenze tra il campione normativo e il campione clinico è stato utilizzato il test t-student. Risultati. Dall'analisi dei dati emerge una buona capacità discriminante del CBA-Y nel distinguere la popolazione normale da quella clinica. Studio 4: Differenze tra i dati normativi e un campione di studenti francesi. Obiettivo. Questo studio vuole indagare come una popolazione diversa da quella italiana risponda al CBA-BG per indagarne gli aspetti di validità esterna. Metodo. La versione italiana del questionario (versione originale) è stata tradotta in francese e successivamente riportata nella lingua originaria (back traslation) in accordo con quanto suggerito nell'articolo di Brislin del 1970. Il questionario in lingua francese è stato somministrato all'interno della facoltà di psicologia di Nanterre (Paris Ouest) durante le pause tra le lezioni. Ogni studente ha compilato il questionario individualmente ed è stato preventivamente istruito a chiedere eventuali chiarimenti all'esaminatore. Hanno partecipato all'indagine 109 studenti (50 maschi; 45,9%) francesi di età compresa tra i 18 e i 24 anni. L'età media dei ragazzi era di 21,98 anni (ds=2,44) e quella delle ragazze era di 20,81 anni (sd=1,51). L'età tra i due generi si differenziava significativamente (F(1,107)=8,35; p<0,01). Tutti gli intervistati erano studenti universitari della facoltà di psicologia di Nanterre. Non vi è differenza statisticamente significativa tra il numero di ragazzi e di ragazze (χ2(1)=0.743; p=0.389). Analisi: per confermare la struttura fattoriale all'interno del campione francese sono stati utilizzati dei modelli di equazioni strutturali nella loro forma più semplice, l'analisi fattoriale confermativa (CFA). A causa delle scarsità del campione ogni scala del CBA-BG è stata valutata singolarmente in modo da ridurre la complessità del modello. Per indagare se vi fossero differenze tra le risposte del campione francese rispetto a quelle di un sottocampione appaiato estratto dai dati normativi è stata utilizzata l'analisi della varianza univariata utilizzando come fattori il genere e il gruppo (francesi e italiani). I risultati dell'analisi presi in esame sono l'interazione tra i fattori e l'effetto principale del gruppo. La prima esplicita la presenza di differenze tra le differenze di genere nei due gruppi mentre il secondo indaga l'eventuale differenza tra i gruppi. L'effetto principale del genere è irrilevante ai fini della presente ricerca. Risultati. L'analisi sul campione francese ha confermato la bontà del questionario per sette delle dieci scale analizzate. La soltanto parziale aderenza del modello ai dati originali è da attribuire alla natura stessa delle scale del CBA-BG. Esse non sono state ricavate tramite procedimenti statistici ma sono state pensate a livello teorico per indagare le differenti aree che in fase di stesura dello strumento erano state considerate utili per la pratica clinica. I due campioni presi in esame erano equivalenti sotto tutti gli aspetti salienti. Oltre che l'appaiamento per genere, età, scolarizzazione e condizione professionale essi non differivano nemmeno per le condizioni di somministrazione. Una variabile difficile da controllare in sede di analisi a cui spesso non si presta particolare attenzione durante la raccolta dei dati. L'analisi dei punteggi ottenuti nei due campioni non ha fatto emergere differenze sostanziali tra le scale del questionario. Solo la scala di benessere e il numero di item utilizzati per descriversi mostrano un punteggio significativamente minore nel campione francese rispetto a quello italiano. Possiamo concludere dicendo che i risultati di questo studio teso a generalizzare i dati ottenuti nella prima fase di validazione del questionario sono globalmente soddisfacenti.

Collaboration between therapist and client has been put forward as a core element of successful therapeutic encounters. There has been debate as to the nature of collaboration in cognitive behavioural therapy. In the UK this debate has intensified since the introduction of Increasing Access to Psychological Therapies (IAPT) in 2008 as CBT is the favoured therapeutic modality within IAPT. Collaboration in CBT has been conceptualised in dichotomous ways. From one perspective it is constructed in positivistic terms, in which the therapist implements manualised protocols with little consideration for the therapeutic relationship; from the other perspective collaboration is constructed in dialogic terms, in which therapist and client use CBT interventions to consider new meanings that the client deems to be relevant. The current study used a discourse analytic methodology to investigate how CBT therapists construct collaboration in their therapeutic practices. The aim was to explicate interpretive repertories that participants used in the construction of collaboration. Semi-structured interviews were used with 8 CBT therapists. Questions related to the arguments for and against the nature of collaboration in CBT. A client choice interpretive repertoire was used by all participants. It was constructed in various ways in line with either positivistic or dialogic perspectives or elements of both. Individual participants constructed client choice from both perspectives suggesting that the dichotomy in perspectives on collaboration in CBT may not be clear-cut. There is an implication for counselling psychology practitioners to reflect on their use of dichotomous perspectives to conceptualise their professional identities.

Este trabalho tem por objetivo desenvolver o processo de retificação a alta velocidade de superligas utilizando rebolos de CBN vitrificados. Para tanto, foram estudadas diversas condições de dressagem, ou seja, as influências dos parâmetros de dressagem no desempenho do rebolo CBN de liga vitrificada. A partir da escolha desses parâmetros, foi também determinado o desgaste do rebolo em diversas condições de velocidade de avanço, e a qualidade superficial das peças retificadas. Foi utilizado um sistema de mapeamento topográfico do rebolo, baseado na emissão acústica emanada da interação entre rebolo-peça, para o monitoramento do processo. Como resultado foi determinada a influência da velocidade de dressagem e da relação de velocidades na rugosidade da peça retificada. Ficou determinado também, o desgaste do rebolo de CBN em função do volume de material retificado para os diferentes materiais das superligas testadas. Através do sistema de mapeamento topográfico, determinou-se os padrões gráficos do comportamento do rebolo de CBN vitrificado. Sendo assim, verificou-se que o sucesso da aplicação de rebolos CBN vitrificados na retificação de superligas depende da qualidade superficial do rebolo após a dressagem e da escolha certa dos parâmetros de retificação, os quais dependem da constituição das ligas testadas.
The aim of this work is to develop a high speed grinding process for super alloys using vitrified CBN wheels. For that, dressing conditions were studied in order to determine its influence on the CBN wheel performance. From the chosen parameters, the wheel wear and the ground surface quality were measured for different infeed speeds. A topographical mapping system based on acoustic emission was used to monitor the grinding process. As a result, the best dressing speed and speed ratio which generated the smallest surface roughness values could be found. The CBN wheel wear was measured and its ratio to the ground volume was evaluated. Through the mapping system, some visual patterns were obtained defining the grinding wheel behavior when dressing and grinding. Thus, the success vitrified CBN wheel application for super alloys depends basically on the wheel topography after dressing and the grinding parameters, which are dose related to the work material composition.

This paper describes a case based reasoning system that is used to guide the parameters of a segmentation algorithm. Instead of using a fixed set of parameters that gives the best average result over all images, the parameteres are tuned to maximize the score for each image separately. The system's foundation is a set of 20 cases that each contains one 3D MRI image and the parameters needed for its optimal segmentation. When a new image is presented to the system a new case is generated and compared to the other cases based on image similarity. The parameters from the best matching case are then used to segment the new image. The key issue is the use of an iterative approach that lets the system adapt the parameters to suit the new image better, if necessary. Each iteration contains a segmentation and a revision of the result, and this is done until the system approves the result. The revision is based on metadata stored in each case to see if the result has the expected properties as defined by the case. The results show that combining case based reasoning and segmentation can be applied within image processing. This is valid for choosing a good set of starting parameters, and also for using case specific knowledge to guide their adaption. A set of challenges for future research is identified and discussed at length.

Det stora behovet av att kunna prognostisera och beskriva framtida dödligheter på ett bra sätt och svårigheten i detta har lett till att många olika modeller tagits fram. En av dessa modeller är den s.k. CBD modellen. I detta arbete så har denna kalibrerats efter svensk dödlighetsdata och dess lämplighet att användas har undersökts. Inga större problem med modellen fanns, förutom att den inte var så lämplig för de allra äldsta individerna (över 95 år). Modellen jämfördes med SCB:s framtidsprognos, en jämförelse som inte gav så mycket men som antydde att dödlighet av de yngre av 65-95 åringarna skulle gå snabbare enligt CBD modellen än vad SCB förutspått, motsatsen verkade inträffa för de högre åldrarna. De framtida dödligheter som tagits fram med CBD modellen användes sedan för att räkna ut nuvärde på en försäkrings framtida utbetalningar. Övre gränser för dessa utbetalningar räknades ut med konfidensintervall, men även enligt standardmetod enligt Solvens 2. Resultatet visade att den övre gränsen framtagen m.h.a. konfidensgräns låg högre upp än då motsvarande gräns räknats ut enligt Solvens 2 standardmetoden.

Cost Benefit Analysis (CBA) has for a long time been used in transport planning, but it is often questioned. One main argument against CBA is that the results depend largely on assumptions regarding one or a few input factors, as for example the future fuel price or valuation of CO2 emissions. The three papers included in this thesis investigate some aspects of uncertainty in transport CBA calculations. The two first papers explore how changes in input data assumptions affect the CBA ranking of six rail and road investments in Stockholm. The first paper deals with the effect of different land-use assumptions while the second deals with the influence of economic growth, driving cost and public transport fare. The third paper investigates how alternative formulations of the public transport mode choice and route choice affect travel flows, ticket revenues and consumer surplus. These are important factors previously known to affect CBA results. The findings of the first two papers suggest that CBA results are robust concerning different land-use scenarios and single input factors. No change in rank between a road and a rail object is observed in the performed model calculations, and only one change between two road objects. The fact that CBA results seem robust regarding input assumptions supports the use CBA as a tool for selecting transport investments. The results in the third paper indicate that if there is detailed interest in, for example, number of boardings and ticket income from a certain transit line, or the total benefit of a price change, a more detailed formulation of the public transport mode choice and route choice will provide more reliable results. On the other hand, this formulation requires substantially more data on the transit line and price structure than the conventional formulation used in Swedish transport planning, especially in areas with many different pricing systems.

QC 20150414

Les courriers électroniques sont devenus au cours des dernières années un moyen de communication privilégié. Leur nombre atteint des niveaux si élevés que leur manipulation par les entreprises devient difficile manuellement et requiert des méthodes automatiques. Les systèmes de suivi automatique permettent un traitement des messages et une augmentation de la facilité pour les utilisateurs qui désirent créer un courriel en réponse à une requête, à partir de messages archivés dans la bibliothèque de l’entreprise. Un des problèmes majeurs dans ce type d’application est la sélection de courriers électroniques archivés pertinents à la requête nouvellement soumise : il est nécessaire que le texte retourné par le système qui servira de base à la rédaction d’une réponse, soit en corrélation avec le sujet de la question posée. La recherche de la similarité entre les textes est donc une des principales tâches de ce système. L’objectif de nos travaux se situe toutefois à un niveau supérieur à la simple recherche de similarité entre courriers électroniques; nous avons comme objectif d’évaluer la similarité entre des textes comportant peu de mots. Pour cela, notre étude comporte deux parties distinctes: la recherche de similarités entre des mots afin d’élargir le vocabulaire d’un texte, e le calcul de la similarité entre les documents. Notre question de recherche consiste à identifier les techniques les plus pertinentes pour évaluer la similarité textuelle, et déterminer s’il est possible d’améliorer ces techniques par des combinaisons de méthodes de calcul de la similarité sémantique et de détection des cooccurrences de mots. Lors de nos expérimentations, nous avons cherché à améliorer la similarité sémantique entre les mots. Nous avons aussi proposé plusieurs méthodes pour élargir le vocabulaire des documents basées sur les résultats de la première partie de nos travaux. Nous avons enfin mesuré la similarité sémantique entre les documents modifiés, afin de les utiliser comme base à la rédaction d’une réponse à la nouvelle requête. Notre bilan nous indique que, grâce à nos calculs, il est possible d’améliorer les résultats de calcul de la similarité entre les mots (cosinus) en filtrant les cooccurrences. Cependant, une méthode de filtrage ne garantit pas nécessairement des améliorations, et peut même dégrader la capacité de détection de similarité textuelle. Une comparaison avec la technique du tf*idf nous a permis de constater que les résultats améliorés du cosinus, que nous avons obtenus, égalent pratiquement cette technique, sans pour autant la dépasser.
E-mails have recently become a popular mean of communication for exchanges between companies and their customers. However the increasing volume of messages makes manual processing difficult to achieve and automatic methods are foreseen as a more efficient solution. Automatic management systems help users in the processing of the messages and in the creation of a response from the messages kept in the company databases. One important question in this type of application is how to select existing e-mails to respond to a new request. The creation of new response messages requires texts pertaining to the new request topics. Finding similarity between documents is also an important task. Our goal for this research effort was to study how to detect similarity between small documents. To accomplish it, we followed a two-pronged approach: - finding similarity between words in order to augment a document’s vocabulary; - estimating similarity between documents, using all the similar words resulting from the previous step. We dedicated our work to determine the most interesting techniques to detect textual similarity between documents, and to improve those techniques using cooccurrences detection and lexical semantic similarity. During our experimentations, we tried different combinations, using cooccurrences detection and lexical similarity. We proposed techniques to augment the vocabulary of each message, based on different kind of reasoning to improve the estimation of similarity between documents. Our results indicate that the proposed augmentation techniques improve significantly the estimation of document similarity. The best results were obtained when using a combination of cooccurrences filter and cosine metric. However our experiments clearly indicate these results do not overcome the performance of similarity techniques based on tf*idf weights.

Bioactive compounds from Cannabis sativa have been used for millennia to alleviate the symptoms of a range of diseases. The physiological basis of effects such as analgesia, stimulation of hunger and reduction of inflammation was established in the late 20th century with the discovery of cannabinoid receptors but efforts to synthesise safe and potent drugs targeting these proteins have so far failed. The major barrier to research in this area is the instability of the receptors outside of biological settings, rendering elucidation of the binding sites by traditional means difficult. Certain small molecules can interact with the cannabinoid type 1 receptor (CB1) at locations distinct to the primary ligand docking site. Such allosteric modulation of the endocannabinoid system offers significant advantages over using orthosteric drugs and in this research a range of indole based structures were synthesised and tested in an attempt to improve the activity and drug-like nature of a lead compound. A partial structure-activity relationship was established, including the description of the most potent allosteric enhancer of CB1 so far reported. Efforts were also undertaken to investigate the allosteric binding environments using photoactivatable ligands based on a CB1 inhibitor. In combination with mutation studies and computer modelling this technique could allow the rational design of allosteric modulators, a task which is not trivial at present. Two photoactivatable compounds were synthesised and shown to interact with the receptor, with a method for isolating covalently labelled peptide fragments from other biomolecules demonstrated using “click chemistry” and a modified Wang resin. This work may find application in future investigations aiming to produce allosteric pharmaceuticals targeting CB1. Furthermore, the techniques described may be applied to study the binding site of a recently described allosteric endocannabinoid or could potentially be adapted to look at secondary binding domains in other G protein-coupled receptors.

Orientador: Mauro Monteiro Garcia de Carvalho
Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Fisica Gleb Wataghin
Made available in DSpace on 2018-07-21T20:30:43Z (GMT). No. of bitstreams: 1 Sato_JulioNoboru_M.pdf: 1907861 bytes, checksum: 9da572e0cf8e9f1e542ef7f2f309d976 (MD5) Previous issue date: 1996
Resumo: Apresentamos neste trabalho um estudo de crescimento homoepitaxial de GaAs que tem por objetivo calibrar o sistema CBE montado na Unicamp, auxiliando no crescimento de ligas ternárias e quaternárias. A escolha do crescimento de GaAs está baseada na sua grande importância em micro e optoeletrônica, assim como no fato de existir uma ampla referência bibliográfica sobre esta liga binária. Em nosso trabalho, mostramos que o modelo de Robertson e Donnelly pode ser usado para explicar o comportamento das taxas de crescimento obtidas. Mostramos, também, que a troca de marca do craqueador bem como a mudança da temperatura de craqueamento, influenciaram as características morfológicas, elétricas e de raio-X das amostras crescidas. Por fim, verificamos que as dopagens com silício e berílio, para as concentrações estudadas, são proporcionais a taxa de evaporação dos respectivos dopantes
Abstract: In this work we present a study on the growth of homoepitaxy of GaAs. Its goal is to calibrate the Chemical Beam Epitaxy system installed at this University, to help in the growth of ternary and quaternary alloys. The choice for GaAs growth was based on its importance for micro and optoelectronics, as well as on the existence of considerable amount of literature on this binary material. In our work we show that the model proposed by Robertson and Donnelly can be used to explain the behavior of the growth rate with respect to growth parameters. We also show that the use of two different cracker cells affected the morphological, electrical and crystallographic characteristics of the material. These characteristics were investigated as a function of the cracker cell temperature. Finally, we verified that Si and Be doping, for the doping levels considered in this study, are proportional to the evaporation rate of the sources used.
Mestrado
Física
Mestre em Física

Hydropower is a fundamental pillar in the Swedish energy system and accounts for a large part of the Swedish electricity production. The regulation power is also essential for balancing the grid load.  Fortum is one of the leading energy companies within hydropower and Klarälven is a river with high importance, where Fortum owns and operates nine powerplants. These power plants constitute a barrier for the wild salmonids in Klarälven, which need to migrate upstream to reach their spawning area. Since the 1930s, the spawning salmonids have been trapped and transported upstream by lorry. After the spawning period, the smolts and kelts, i.e. the juvenile and spawned salmonids, have to migrate downstream. Due to the lack of fishways, they are forced to pass the eight remaining power plants. This, together with predation, entails a high mortality rate. Two independently performed studies indicates on survival rates of 16 % and 30 %. To stabilize the wild salmonid population, the downstream survival must increase, and a proposed solution is to implement a downstream trap-and-transport solution. This trap implementation could be a step towards the environmental adaptation of hydropower and a part of the action plan proposed in June 2016, during the Agreement on Swedish energy policy. To find the most cost-beneficial environmental measure, a socio-economic assessment method can be used. A Cost-Benefit Analysis (CBA) is a commonly used socio-economic method, which evaluates the benefits and costs during the entire project lifetime. Energiforsk has, within the project FRAM-KLIV, developed a CBA tool that aims to simplify the socio-economic evaluation. The purpose of this thesis was to investigate whether the CBA tool can be used in future permit processes to prioritize between different project proposals to find the most beneficial environmental improvements. In order to evaluate the tool, it was to be applied on the trap-and-transport project in Edsforsen to evaluate the possibility of a socio-economic profitability. Also, the concepts and theory behind CBA was to be analysed and the suitability of translating environmental consequences into monetary values was to be evaluated. In the analysis of Edsforsen, 13 scenarios were developed. The first scenario served as a basis for the other scenarios, which were created as a sensitivity analysis. The result of the CBA showed a large socio-economic benefit and the most important parameter was identified as people’s willingness to pay for an increase of the wild salmonid stock in Klarälven. In the CBA, this parameter had a high uncertainty, as it was based on a survey performed for another project in another part of Sweden. It was found that in order for the socio-economic result to be positive, all households in Sweden must be willing to pay at least 35 SEK. As a complement to the CBA result, an evaluation regarding the marginal cost per fish was performed and an interval of 50-580 SEK per smolt was obtained. However, the calculations were based on several uncertainties and the interval should therefore be interpreted as a guideline rather than a precise result. It was concluded that in situations when a socio-economic analysis is required, and when it is possible to express consequences in monetary values, the method of CBA is appropriate. It is also a suitable methodology for evaluations of large projects, as it provides a comprehensible overview of the costs and the benefits. Despite the criticism directed towards CBA regarding uncertainties and its anthropogenic perspective, it could be concluded that using CBA as a socio-economic assessment method provides a perspicuous and quantitative result. Thus, the usage of CBA in prioritization processes of different environmental measures can be highly useful. Energiforsk’s CBA tool provides a framework with guidelines that can be highly useful and accelerate the analysis process. However, the performance of the tool version used in this thesis was not fully satisfactory due to a few malfunctions. The tool is still under development and it is likely that these errors will be adjusted in future versions. If the malfunctions in the tool would be adjusted, it could become useful for authorities, companies and other actors that wants to evaluate hydropower related environmental measures or when prioritizing between different project proposals to find the most beneficial environmental improvements in future permit processes.
Vattenkraften är en grundläggande pelare i det svenska energisystemet och står för en stor andel av den svenska elproduktionen på årsbasis. Vattenkraften har även hög betydelse för det svenska nätets balanseringsförmåga. Fortum är ett av de ledande energibolagen inom vattenkraft i Sverige och en viktig älv för dess kraftproduktion är Klarälven. Denna älv är dessutom fundamental för Klarälvslaxen och Klarälvsöringen och dess nödvändiga lekvandring. Under naturliga förhållanden ska den lekmogna laxen vandra uppströms Klarälven, men denna vandring försvåras av vattenkraftverken. Sedan 1930-talet har laxen transporterats uppströms Klarälven med lastbil för att nå sina lekområden, men när smolten och kelten ska vandra nedåt för att nå Vänern på nytt måste den passera de resterande åtta vattenkraftverken, vilket tillsammans med predation resulterar i en hög dödlighet. Två oberoende studier indikerar en överlevnad på 16 % respektive 30 %. För att stabilisera det vilda laxbeståndet föreslås en uppfångsanordning med nedströmstransport, där fisken fångas upp i en fälla och transporteras nedströms med lastbil. I juni 2016 beslutades det genom Energiöverenskommelsen att vattenkraften ska miljöanpassas och ett genomförande av en sådan fälla skulle möjligen kunna vara en sådan miljöanpassning. För att finna de mest lönsamma miljöanpassningarna inom vattenkraften bör samhällsekonomiska lönsamhetsbedömningar genomföras. En kostnadsnyttoanalys (CBA) är sådan bedömningsmetod där kostnader och nyttor genererade under projektets livstid ställs mot varandra. Genom projektet FRAM-KLIV har Energiforsk varit med och tagit fram ett CBA-verktyg som ska underlätta genomförandet av en sådan beräkningsanalys. CBA-verktyget användes som ram i det här examensarbetet där en kostnadsnyttoanalys gällande laxens förbättrade nedströmspassage genomfördes och utvärderades. Syftet med detta examensarbete har varit att undersöka huruvida detta CBA-verktyg skulle kunna användas i framtida prioriteringar under tillståndsprocesser för att hitta de mest lönsamma miljöanpassningarna. Dessutom skulle bakomliggande koncept och teori analyseras. Tillika skulle en utvärdering gällande lämpligheten i att värdera miljövärden i monetära termer utföras. I arbetet utformades totalt tretton scenarier, varav det första ansågs som grundscenariot och utgjorde basen för de övriga tolv, vilka skapades för att identifiera de mest kritiska parametrarna genom en känslighetsanalys. Kostnadsnyttoanalysens erhållna resultat tydde på en stor samhällsekonomisk lönsamhet och den mest kritiska parametern identifierades som människors betalningsvilja för en ökad vildlaxpopulation i Klarälven. Osäkerheten kring denna parameter var stor då den baserats på en annan projektundersökning i en annan del av landet. För att erhålla ett positivt samhällsekonomiskt resultat visade denna undersökning att det måste finnas en betalningsvilja på minst 35 kr per hushåll i Sverige. Som ett komplement till resultatet utfördes en beräkning gällande hur mycket varje smolt skulle behöva vara värd för att den samhällsekonomiska nyttan ska vara lika stor som kostnaden.  Beräkningen gav ett intervall mellan 50 och 580 kr, men då använd data innehöll stora osäkerheter bör resultatet ses mer som en riktlinje och inte ett exakt resultat. En slutsats som kunde dras var att det anses lämpligt att uttrycka miljövärden i monetära värden när en samhällsekonomisk analys är nödvändig. CBA som metod anses också vara lämplig och användbar när större projekt ska analyseras eftersom den ger en bred bild över fördelningen mellan kostnader och nyttor. Metoden förser dessutom användaren med ett lättolkat och överskådligt resultat och därav skulle CBA som metod vara lämplig gällande prioritering av miljönyttor. Energiforsks CBA-verktyg kan underlätta processen i och med att det ger användaren en ram med riktlinjer för hur beräkningsprocessen ska gå till. Verktygsversionen som användes i detta examensarbete uppfyllde dock inte full tillfredsställelse vad gällde dess funktionalitet. Verktyget är visserligen fortfarande under utveckling och det är möjligt att dessa funktionsfel kommer att justeras i framtida versioner. Om dessa justeringar sker kan verktyget komma att bli fullt funktionsdugligt och vara användbart för myndigheter, företag och andra aktörer som vill utvärdera vattenkraftrelaterade miljöåtgärder eller som prioriteringsverktyg gällande miljöförbättringar inom framtida tillståndsprocesser.

Physiology
Ph.D.
c-Cbl protein functions as an E3 ligase and scaffolding protein, where three residues, Y700, Y731, and Y774, upon phosphorylation, have been shown to initiate several signaling cascades. In this study, we investigated the role of these phospho-tyrosine residues in the platelet functional responses upon integrin engagement. We observed that c-Cbl Y700, Y731 and Y774 undergo phosphorylation upon platelet adhesion to immobilized fibrinogen, which was inhibited in the presence of PP2, a pan-src family kinase (SFK) inhibitor, suggesting that c-Cbl is phosphorylated downstream of SFKs. However, OXSI-2, a Syk inhibitor, significantly reduced c-Cbl phosphorylation at residues Y774 and Y700, without affecting Y731 phosphorylation. Interestingly, PP2 inhibited both platelet spreading on fibrinogen as well as clot retraction, whereas OXSI-2 blocked only platelet spreading, suggesting a differential role of these tyrosine residues. The physiological role of c-Cbl and Y731 was studied using platelets from c-Cbl KO and c-CblYF/YF knock-in mice. c-Cbl KO and c-Cbl YF/YF platelets had a significantly reduced spreading over immobilized fibrinogen. Furthermore, clot retraction with c-Cbl KO and c-Cbl YF/YF platelets was drastically delayed. These results indicate that c-Cbl and particularly its phosphorylated residue Y731 plays an important role in platelet outside-in signaling contributing to platelet spreading and clot retraction
Temple University--Theses

Le complexe de liaison de la coiffe des ARN (CBC) joue un rôle essentiel dans leur maturation et déclenche une variété de réactions biochimiques, via son interaction avec différents partenaires. Deux complexes, CBC-ARS2-PHAX (CBCAP), et CBC-ARS2-ZC3H18-NEXT (CBCN), ont récemment été montré comme important pour cibler les ARN vers l'export (CBCAP) ou la dégradation (CBCN). Cependant, les mécanismes par lesquels la sélection se fait pour l'une voie ou l'autre reste mystérieuse. Ainsi, une question majeure qui reste à résoudre est de savoir quand et comment ces complexes sont recrutés sur les ARN. Dans ce travail, j'ai utilisé la procédure du iCLIP (Cross-Linking and Immuno-Precipitation), afin d'identifier les cibles de ces complexes sur l'ensemble du transcriptome humain. J'ai réalisé un iCLIP sur cinq composants de CBCAP et CBCN, et j'ai comparé les résultats à ceux obtenus avec RBM7, un composant de NEXT précédemment étudié par iCLIP. Mes résultats indiquent que: (i) CBP20, ARS2, PHAX et ZC3H18 se lient près de la coiffe des ARN, tandis que RBM7 et MTR4 se lient partout; (ii) CBP20, ARS2, PHAX et ZC3H18 s'associent à un large ensemble d'ARN transcrits par l'ARN polymérase II et montrent une faible sélectivité; (iii) la liaison de ces protéines varie avec l'état de maturation des ARN, avec le CBC enrichi sur les ARN matures, tout comme ARS2/PHAX/ZC3H18 et MTR4 (bien que dans une moindre mesure), tandis que RBM7 est préférentiellement lié sur les pre-mRNAs non épissés; (iv) une liaison différentielle de RBM7 et MTR4 sur les ARN, avec RBM7 enrichi sur les introns et les PROMPTs, et MTR4 plus présent sur les ARN mature. Bien que des expériences additionnelles soient requises, nous proposons que le CBCAP et le CBCN se lient à un même ensemble d'ARN, ce qui indique à la fois une compétition entre ZC3H18 et PHAX pour la liaison à ces ARN, et l'absence de voies de routage bien déterminées qui ciblerait les ARN vers l'une ou l'autre de ces protéines. Le devenir des ARN pourrait ainsi être déterminé par d'autres caractéristiques des ARN, ou encore par des protéines additionnelles. Ces facteurs pourraient s'allier aux protéines liées à la coiffe afin de favoriser la formation du CBCAP ou du CBCN. Dans le but d’identifier des facteurs additionnels, j'ai réalisé un screen d'interaction par spectrométrie de masse après purification de ARS2 ou CBP80. Ceci a été fait dans des conditions natives ou après un cross-link des complexes à la formaldéhyde, afin de stabiliser les interactions transitoires. Ceci a permis d'identifier de nouveaux partenaires de ARS2 et de CBP80, dont la majorité sont impliqués dans l'épissage des ARN. Des expériences additionnelles seront nécessaires pour valider ces interactions
The cap-binding complex (CBC) plays a pivotal role in post-transcriptional processing events and orchestrates a variety of metabolic pathways, through association with different interaction partners. Two CBC sub-complexes, the CBC-ARS2-PHAX (CBCAP) and the CBC-nuclear exosome targeting (NEXT) complex (CBCN), were recently shown to target capped RNA either toward export or degradation, but the mechanisms by which they can discriminate between different RNA families and route them toward different metabolic pathways still remain unclear. A major question to be answered is how and when the different CBC subcomplexes are recruited to the RNP. Here, we used an individual nucleotide-resolution UV cross-linking and immunoprecipitation (iCLIP) approach to identify the transcriptome-wide targets for 5 different components of the CBCAP and CBCN complexes, and compared results to the previously analysed NEXT-component RBM7. We report that: (i) CBP20, ARS2, PHAX and ZC3H18 bind close to the cap, while RBM7 and MTR4 bind throughout the mRNA body; (ii) CBP20, ARS2, PHAX and ZC3H18 associate with a broad set of RNA polymerase II (PolII)-derived RNAs and have only mild species preferences; (iii) binding varies with the RNA maturation stage, with the CBC being highly enriched on mature mRNA, ARS2/PHAX/ZC3H18/MTR4 less so, and RMB7 preferentially bound to pre-mRNAs; (iv) MTR4 and RBM7 show different specificities, with RBM7 being highly enriched on introns and promoter upstream transcripts (PROMPTs), while MTR4 is additionally present on mature RNAs. Although more experimental work is needed to fully support our model, we propose that CBCAP and CBCN bind overlapping sets of RNAs, indicating a competition between the proteins ZC3H18 and PHAX, and the lack of a strict RNA sorting mechanism. RNA fate may therefore be determined by additional RNA features and/or by other RNA-binding proteins, which may synergize with the cap and drive the formation of one specific CBC subcomplex instead of another. In an attempt to identify yet unknown factors that may interact with cap-bound CBCAP and CBCN, we performed a protein interaction screen leveraging affinity capture-mass spectrometry (ACMS), using ARS2 and CBP80 as bait proteins. As a complementary approach, we also employed a formaldehyde-based chemical cross-linking strategy, aimed at stabilizing weak/transient interactions. Although we failed to detect any transient interactions involving the CBC, we identified several potential CBC80 and ARS2 interactors, the majority of which are involved in pre-mRNA splicing. Additional quantitative experiments are required to validate our ACMS results and confirm the existence of such protein interactions in vivo

The Cbl family of E3 ubiquitin ligases are important negative regulators of multiple receptor and cytoplasmic tyrosine kinases, and participate in a wide variety of cellular processes. Uncoupling of Cbl-mediated negative regulation allows activated receptor tyrosine kinases such as the Met receptor to escape degradation, enhancing their oncogenic potential in vitro and in vivo. Despite the consequences of loss of Cbl-mediated negative regulation for human disease, little is known about the mechanisms regulating Cbl protein levels themselves.
In this thesis work, I demonstrate a differential regulation of c-Cbl and Cbl-b downstream of the Met receptor tyrosine kinase. Cbl-b protein levels decrease in response to Met kinase activity, whereas c-Cbl levels remain stable. Cbl-b is partially degraded in a proteasome-dependant manner. This requires Cbl-b ubiquitin ligase activity and a carboxy terminal domain region located between the RING and UBA domains. I conclude that the regulation of c-Cbl and Cbl-b differs downstream of Met, and propose that negative regulation of Cbl-b by a dysregulated Met receptor may contribute to tumourigenesis.

La trisomie 21 est une maladie génétique caractérisée par la présence de trois chromosomes 21 au lieu de deux dans le génome. Ce phénomène ne provoque pas la synthèse de protéines anormales, mais est responsable de perturbations biochimiques quantitatives et d’anomalies métaboliques. Une de ces enzymes, la Cystathionine b-Synthase (EC 4. 2. 1. 22, CBS), nous a particulièrement intéressée. La surexpression de la CBS perturbe directement le métabolisme de l’homocystéine et a également pour conséquence une surproduction d’H2S. H2S, nouvellement connu pour être un neuromodulateur à des concentrations physiologiques, est un gaz toxique. Une surproduction de ce dernier a été montrée dans le cerveau des personnes atteintes de trisomie 21. Au vu de ces informations, l’hypothèse suivante a été avancée : l’inhibition réversible de la surexpression de la CBS limiterait les désordres métaboliques à l’origine du développement du retard mental. Ce travail a consisté à réaliser des tests enzymatiques in vitro sensibles pour déceler, au sein de chimiothèques, la présence de molécules possédant un effet inhibiteur sur la cystathionine β-synthase. Plus de 7000 molécules provenant de différentes chimiothèques ont ainsi été criblées. 8 molécules appartenant à 3 familles structurales se sont avérées capables d’inhiber réversiblement et spécifiquement l’activité enzymatique de la CBS humaine recombinante et la CBS extraite de foie de Rat. Nous avons alors pu déterminé à l’aide de courbes effet-doses les CI50 et les possibles sites d'action des molécules sur l'enzyme. Dans un deuxième temps, des pharmacomodulations ont été réalisées pour chaque famille de molécules (coumarine, pyridazine et benzothiophène) pour tenter d'améliorer le pouvoir inhibiteur des « hits »
Down syndrome has a genetic basis: the presence in the genome of three rather than two chromosomes 21. This leads to gene dosage effects: involved genes and gene products are normal but their productions are increased. One enzyme, the Cystathionine b-Synthase (EC 4. 2. 1. 22, CBS) particularly interested us. The over expression of CBS directly disturbs the homocysteine metabolism and has special consequence: an overproduction of H2S. H2S is a well-known toxic gas but it is recently known as a neuromodulator in physiological concentrations. An overproduction of H2S was shown in the DS people’s brain. In view of all this information, the following hypothesis was advanced: a reversible inhibition of CBS activity could limit the metabolic disorders which are the origin of development of the intellectual retardation. This work consisted in carrying out significant in vitro enzymatic tests to detect, within libraries of chemicals, the presence of molecules having an inhibiting effect on the cystathionine β-synthase. More than 7000 molecules from different chemical libraries have been screened. 8 molecules belonging to three chemicals structures were able to inhibit reversibly and specifically the two enzymatic activities assays: pure recombinant human CBS and extracts from rat livers. We determined by means of dose-response curve theirs IC50 and the possible action site of molecules on the enzyme. In a second time, pharmacomodulations were realized for each family of molecules (coumarine, pyridazine and benzothiophen) in the purpose of improving the inhibitive power of « hits »

Introduction: This thesis took a successfully trialed, fully automated Internet based CBT (iCBT) program for treating socially anxious stuttering adults, known as CBTpsych (see Helgadottir, Menzies, Onslow, Packman & O'Brian, 2009b), and adapted it for stuttering adolescents. This saw the development and phase I trial of a fully automated iCBT program that targets social anxiety in stuttering adolescents, and does not require clinician involvement. Method: The adolescent iCBT program used algorithms to tailor treatment to each user’s specific issues relating to social anxiety. All seven modules in the iCBT program were designed around Clark and Wells’ (1995) model of social anxiety. Twelve stuttering adolescents presenting for stuttering treatment at the Australian Stuttering Research Centre (ASRC) were offered iCBT program access for five months. Participants were not allowed access to a psychologist during the trial. Results: Of the 12 participants enrolled in the study, three never started the iCBT program. Nine participants used the iCBT program, and 44% (n = 4/9) completed it. Only four participants returned their post-treatment assessment booklets. Post-treatment analysis revealed reductions in assessment scores for those who completed the program. Post-trial phone interviews with five of the nine participants who used the program suggested: (1) a positive response to iCBT; (2) that time constraints and school pressures act as a serious barrier for this age group accessing Internet based treatment. Conclusion: In line with earlier findings on this age group, stuttering adolescents were shown to be a difficult population to engage, evident by the high non-completion rate of the iCBT program (42%, n = 5/12) and the high non-completion rate of post-trial assessment booklets (67%, n = 8/12). More research is needed to find ways of lifting adolescent completion rates with Internet based treatment.

In this thesis we describe our study of two knowledge intensive Conversational Case-Based Reasoning (CCBR) systems and their methods. We look in particular at the way they have solved inferencing and question ranking. Then we continue with a description of our own design for a CCBR system, that will help patients share their experiences of side effects with drugs, with other patients. We describe how we create cases, how our question selection methods work and present an example of how the domain model will look. It is also included a simulation of how a dialogue would be for a patient. The design we have created is a good basis for implementing a knowledge intensive CCBR system. The system should work better than a normal CCBR system, because of the inferencing and question ranking methods, which should lessen the cognitive load on the user and require fewer questions answered, to reach a good solution.

Le p300/CBP acétyltransférase est un co-activateur transcriptionnel très important qui est impliqué dans la régulation d'un grand nombre de processus biologiques, comme la transcription d'ADN, le développement et l'immunité innée. Jusqu'à présent, le rôle de p300/CBP dans la régulation de l'expression des gènes a été largement étudiée, mais les mécanismes qui régulent son activité enzymatique sont encore peu connus. Des études ont montré que le dysfonctionnement de p300/CBP est associé à plusieurs formes de cancer et de maladies neurodégénératives. Dés lors, chaque progrès concernant les mécanismes de régulation de p300/CBP est devenu primordial pour le développement de nouvelles thérapies. Le 'noyau' de p300/CBP contient deux domaines pour la reconnaissance des modifications post-traductionnelles (MPTs), un bromodomaine et un PHD finger (le module BP), adjacent à un domaine HAT (ou domaine histone acétyltransférase). Plusieurs enzymes, modifiant la chromatine, contiennent des domaines de reconnaissance des MPTs. Fréquemment des groupements particuliers de ces domaines sont très conservés et liés, au sein de la même protéine ou du même complexe protéique, suggérant qu'ils réalisent des fonctions coordonnées. Ces domaines adjacents peuvent agir en concertation dans la reconnaissance simultanée de différents MPTs ou peuvent exercer des fonctions différentes de celles qui sont effectuées par ces deux domaines particuliers, tels que les fonctions de régulation enzymatique. Plusieurs études suggèrent que les cycles acétylation/désacétylation dans la boucle d'auto-inhibition, à l'intérieur du domaine HAT, jouent un rôle important dans la régulation de l'activité enzymatique de p300/CBP. La proximité du module BP et du domaine HAT suggère que la spécificité de liaison, appartenant au module BP, peut être intrinsèquement liée à la régulation de l'activité du domaine HAT. L'objectif de ma thèse est de déterminer le rôle du module BP dans la régulation de l'activité du domaine HAT. Je propose que le module BP soit impliqué dans la régulation de p300/CBP de deux façons. La première consiste à établir un lien avec le domaine HAT qui stabilise la conformation auto-inhibée de l'enzyme. La deuxième exige que le module BP joue un rôle dans le choix des substrats de p300/CBP. J'ai été en mesure de montrer que BP peut se lier au domaine HAT et à la chromatine modifiée et qu'il peut reconnaître les modifications effectuées par p300/CBP lui-même. Les données obtenues indiquent que le module BP peut être impliqué dans la régulation de l'activité de p300/CBP et dans son ciblage à la chromatine.

This thesis examines CBD (Central Business District) land value variations. The objectives of this study are (1) to compare monocentric and nonmonocentric models and (2) to analyze the determinants of CBD land value variations. Transactions of vacant land from 1975 to 1987 in Central Ward (Chuo Ku), Tokyo, comprise the data base for this study. A monocentric model and nonmonocentric model are compared using a negative exponential function and trend surface analysis (based on a double power series of location coordinates). For the comparison, three-dimensional pictures and contour maps are utilized as well as statistics of goodness-of-fit and predictive powers. To analyze determinants of CBD land value variations, we employ a hedonic-price approach. Trend surface analysis is superior to the monocentric model in terms of goodness-of-fit and predictive powers. However, centrality is still an important determinant. Proximity to subway or railway stations, or to the Ginza shopping area, are also important factors. Other influential determinants include time of sale, lot shape, corner location, road width, and floor area ratio.
Business, Sauder School of
Graduate

Ce projet de maîtrise consiste à l'asservissement et la mise en marche d'un réacteur d'épitaxie par jets chimiques au Laboratoire d'Épitaxie Avancée de l'Université de Sherbrooke. Le réacteur sert à la croissance dans l'ultravide de matériaux semi-conducteurs tels que l'arséniure de gallium (GaAs) et le phosphure d'indium-gallium (GalnP). La programmation LabVIEW™ et du matériel informatique de National Instruments sont utilisés pour asservir le réacteur. Le contrôle de la température de l'échantillon et de la pression de contrôle des réactifs de croissance dans le réacteur est assuré par des boucles de rétroaction. Ainsi, la température de l'échantillon est stabilisée à ±0,4 °C, alors que les pressions de contrôle de gaz peuvent être modulées sur un ordre de grandeur en 2 à 4 secondes, et stabilisées à ±0,002 Torr. Le système de pompage du réacteur a été amélioré suite à des mesures de vitesse de pompage d'une pompe cryogénique. Ces mesures révèlent une dégradation sur plus d'un ordre de grandeur de son pompage d'hydrogène avec l'opération à long terme. Le remplacement de la pompe cryogénique par une pompe turbo-moléculaire comme pompe principale a permis d'améliorer la fiabilité du système de pompage du système sous vide. D'autre part, la conductance du système d'acheminement de gaz et d'injection a été augmentée afin de réduire un effet mémoire des sources le système et faciliter la croissance de matériaux ternaires. Ainsi, des croissances de GaAs (100) sur substrat de même nature ont été effectuées et ont révélé un matériau de bonne qualité. Sa rugosité moyenne de 0,17 nm, mesurée par microscopie à force atomique, est très faible selon la littérature. De plus, une mobilité élevée des porteurs est obtenue à fort dopage au silicium, au tellure et au carbone, notamment une mobilité de 42 ± 9 cm2V_1s_1 des porteurs majoritaires "(trous) lors du dopage au carbone à 1,5 • 1019 cm-3, en accord avec la courbe théorique. La croissance du matériau ternaire GalnP a aussi été réalisée en accord de maille avec le substrat de GaAs, et avec une rugosité de 0,96 nm. Ceci constitue un premier pas dans la croissance d'alliages ternaires au laboratoire. Finalement, la mise eh marche du réacteur d'épitaxie par jets chimiques permet maintenant à cinq étudiants gradués de faire progresser des projets reliés directement à la croissance épitaxiale au Laboratoire d'Épitaxie Avancée de l'Université de Sherbrooke. [symboles non conformes]

Suomen sorateiden kunnon heikkeneminen on ollut polttava puheenaihe jo jonkin aikaa. Yhtenä osana sorateiden kunnossapitoa on selvittää soratien rakennekerrosten kantavuus. Soratien kantavuuden määrittämiseksi tierakenteesta tulee selvittää materiaaliolosuhteet, olosuhdetekijät ja jännitystilaa kuvaavat tekijät. Materiaaliolosuhteisiin vaikuttavia tekijöitä ovat muun muassa maksimiraekoko, rakeisuuskäyrän muoto ja hienoainespitoisuus. Olosuhdetekijöitä kuvaavia tekijöitä ovat muun muassa tiiveystila, routimis-sulamisprosessi ja kosteustila. Jännitystilaa kuvataan kenttä- ja laboratoriotutkimuksilla saatavilla muodonmuutosominaisuuksilla. CBR- ja kolmiaksiaalikoe ovat yleisimmin käytössä olevat laboratoriomenetelmät muodonmuutosominaisuuksien määrittämiseksi. Kokeiden tuloksena saadaan muodonmuutosmoduuli, jota voidaan hyödyntää soratien kantavuutta määritettäessä. Työn tavoitteena on selvittää CBR- ja kolmiaksiaalikokeen eroavaisuuksia ja antaa suosituksia, mitä menetelmää tulisi käyttää missäkin tilanteessa. Työ suoritetaan kirjallisuuskatsauksena, jonka lisäksi työhön kuuluu käytännön osuus, jossa työn tekijä suorittaa CBR- ja rakeisuuskäyräkokeita Oulun Yliopiston geotekniikan laboratoriossa. Kirjallisuuskatsauksen perusteella ei voida antaa tarkkoja rajoja, milloin käyttää mitäkin menetelmää. Sekä CBRettä kolmiaksiaalikokeessa näytteen edustavuudella on suuri merkitys koetuloksiin. Molemmissa kokeissa voidaan joutua poistamaan suuria rakeita näytteen muodostamiseksi, jolloin edustavuus heikkenee. CBR-kokeessa tuloksena saadaan CBR-luku, jota verrataan empiirisesti saatuun standardilukuun. Kolmiaksiaalikokeessa tuloksena saadaan jännitysparametrit, joiden avulla voidaan määrittää näytteen koheesio ja kitkakulma. CBR-kokeessa näytettä puristetaan yksiaksiaalisesti. Kolmiaksiaalikokeessa näytettä puristetaan kolmesta jännityssuunnasta. kolmiaksiaalisessa puristuksessa ongelmaksi muodostuu jännitysten kiertyminen, sillä kolmiaksiaalikokeessa jännityssuuntien kiertymistä ei oteta huomioon
Declining conditions in Finland’s gravel roads have been hot topic for a while. One part of gravel road maintenance is to find out loading capacity of the road. To define loading capacity, material conditions, situational features and stress state needs to be defined. Factors affecting material conditions are among other things maximum grain size, grading curve and fine content. Factors describing situational features are for example tightness, frost-melt cycle and moisture content. Stress state is defined by deformation features in field survey and laboratory tests. CBR- and Triaxial compression tests are most used laboratory tests to determine deformation features. Deformation modulus is received as a result from both tests. Modulus can be used to define loading capacity of gravel road. Objective of this thesis is to find out differences between CBR- and Triaxial compression tests and give recommendations which test to use at given situation. Thesis is performed as a literature review. In addition, author does CBR- and grading curve tests as a practical part in Geotechnical Laboratory of University of Oulu. Based on literature review, no precise boundaries can be given for method selection. Both CBR- and Triaxial compression tests are very dependent on how well specimen represents real conditions. In both tests, large grains may need to be removed to construct specimen. In this case, representativeness of the specimen declines. In CBR-test, the result is a CBR value, which is then compared to standard CBR value. In Triaxial test, the test results are stress parameters, which are used to determine cohesion and angle of friction of specimen. In CBR-test, specimen is compressed uniaxially, where as in Triaxial test specimen is compressed triaxially. Triaxial test doesn’t take into consideration twisting of stresses which becomes a problem

Les récepteurs nucléaires (NRs) forment une famille de facteurs de transcription dont l'activité est contrôlée par la fixation de ligands. La liaison à un ligand agoniste permet le recrutement de coactivateurs transcriptionnels, comme les p300/CREB‑Binding­Protein (p300/CBP) et les p160/Steroid‑Receptor­‑Coactivator (p160/SRC). Ces protéines possèdent plusieurs motifs LxxLL (L = Leucine ; X = n'importe quel acide aminé), responsables de l'interaction aux NRs. Nous avons caractérisé le domaine de CBP interagissant aux LBDs et co‑cristallisé un complexe de l'extrémité Nterminale de CBP (Nter‑CBP) et du LBD du Peroxisome Proliferator‑Activated Receptor gamma (lbd‑PPARg). La mauvaise qualité des cristaux de Nter‑CBP/lbd‑PPARg n'a pas permis de déterminer la structure du complexe. L'étude de Nter‑CBP par résonance magnétique nucléaire (RMN) montre que ce fragment isolé est peu structuré. Son interaction aux LBDs de trois NRs - PPARg, Retinoid X Receptor alpha (RXRa) et Estrogen Related Receptor gamma (ERRg) - a été étudiée par RMN. En raison d'une forte atténuation du signal des résidus impliqués dans l'interaction aux LBDs, la structure du fragment de mCBP en complexe à un LBD n'a pu être résolue, mais la zone d'interaction a été cartographiée : les résidus de CBP impliqués dans l'interaction représentent deux fragments discontinus, délimités par les acides aminés (31 à 38) et (61 à 80), la zone (61 à 80) contenant un LxxLL. L'interaction de fragments de CBP ou de SRCs aux LBDs de PPARg et RXRa a été étudiée de façon comparative par résonance plasmonique de surface (SPR) et par électrophorèse en conditions natives (N‑PAGE). Les résultats montrent l'existence d'interactions préférentielles entre RXRa et les SRCs, et entre PPARg et CBP. Les conclusions de nos travaux suggèrent que CBP puisse être recruté directement par l'hétérodimère PPARg/RXRa, ce qui pourrait jouer un rôle dans la permissivité de cet hétérodimère
Nuclear receptors form a large family of transcription factors, whose activities are often controled by the fixation of small ligands. Binding to an agonist ligand allows the recruitment of transcriptional coactivators, like the p300/CREB‑Binding­Protein (p300/CBP) or the p160/Steroid‑Receptor­‑Coactivator (p160/SRC). Those coactivators possess several LxxLL motifs (L = Leucine ; X = any aminoacid), mainly responsible for the interaction with NRs. We characterized the aminoterminal (Nter) domain of CBP responsible for the interaction with NR Ligand Binding Domains (LBDs) and cocrystallized it together with the Peroxisome Proliferator‑Activated Receptor gamma LBD (lbd‑PPARg). The poor diffraction properties of the crystals did not allow us to solve the structure of this (Nter-CBP/lbd-PPARg) complex. Nuclear Magnetic Resonance (NMR) experiments demonstrate that Nter-CBP is poorly structured on its own. Its interaction with the LBDs of 3 different NRs - PPARg, Retinoid X Receptor alpha (RXRa) and Estrogen Related Receptor gamma (ERRg) – was studied by NMR. The structure of CBP in complex with any of these LBDs could not be solved, because of a strong signal attenuation of the residues implied in the interaction. Nevertheless, the aminoacids of CBP that interact with those 3 NRs could be mapped by this technique : they correspond to 2 discontinuous zones, delimited by the aminoacids (31 to 38) and (61 to 80), the latter containing the LxxLL motif. We also studied the interaction between CBP or SRCs fragments with lbd-PPARg and lbd-RXRa, in a comparative manner, by surface plasmon resonance and native electrophoresis. Our results show that CBP interacts preferentially with PPARg, whereas SRCs show a better affinity towards RXRa. We suggest that CBP might be directly recruited by PPARg/RXRa, which could play a role in the permissivity of this heterodimer